Gene cloning and characterization of VcrM, a Na+-coupled multidrug efflux pump, from Vibrio cholerae non-O1

Nazmul Md Huda, Jing Chen, Yuji Morita, Teruo Kuroda, Tohru Mizushima, Tomofusa Tsuchiya

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Abstract

We cloned a DNA fragment responsible for drug resistance from chromosome of Vibrio cholerae non-O1. Nucleotide sequence analysis of this fragment revealed the presence of a single open reading frame encoding a protein consisting of 445 amino acid residues. We designated the gene as vcrM. Hydropathy analysis of the deduced amino acid sequence of VcrM suggests the presence of 12 trans-membrane segments. A dendrogram showed that VcrM is a member of the DinF-subfamily within the MATE family of multidrug efflux pumps. Expression of the cloned vcrM gene in drug-hypersensitive Escherichia coli KAM32 cells made them resistant to acriflavine, 4′, 6-diamidino-2-phenylindole, Hoechst 33342, rhodamine 6G, tetraphenylphosphonium chloride (TPPCI) and ethidium bromide. Efflux of acriflavine due to VcrM was dependent on Na+ or Li+. Moreover, Na+ efflux was observed with VcrM when TPPCI was added to Na+-loaded cells. Therefore, we conclude that VcrM is a Na+/drug antiporter-type multidrug efflux pump.

Original languageEnglish (US)
Pages (from-to)419-427
Number of pages9
JournalMicrobiology and Immunology
Volume47
Issue number6
DOIs
StatePublished - Jan 1 2003

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Keywords

  • Drug efflux pump
  • Multidrug resistance
  • VcrM
  • Vibrio cholerae non-O1

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Virology

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